CN115404229A - 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 - Google Patents
双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 Download PDFInfo
- Publication number
- CN115404229A CN115404229A CN202210554944.8A CN202210554944A CN115404229A CN 115404229 A CN115404229 A CN 115404229A CN 202210554944 A CN202210554944 A CN 202210554944A CN 115404229 A CN115404229 A CN 115404229A
- Authority
- CN
- China
- Prior art keywords
- terpene synthase
- leu
- compound
- ser
- glu
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010087432 terpene synthase Proteins 0.000 title claims abstract description 95
- 230000001588 bifunctional effect Effects 0.000 title claims abstract description 86
- 230000003197 catalytic effect Effects 0.000 title claims abstract description 42
- 229930002368 sesterterpene Natural products 0.000 title claims abstract description 19
- -1 ring system sesterterpene compound Chemical class 0.000 title abstract description 14
- 150000001875 compounds Chemical class 0.000 claims description 51
- 229940125898 compound 5 Drugs 0.000 claims description 23
- 108020004707 nucleic acids Proteins 0.000 claims description 15
- 102000039446 nucleic acids Human genes 0.000 claims description 15
- 150000007523 nucleic acids Chemical class 0.000 claims description 15
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 12
- 238000003259 recombinant expression Methods 0.000 claims description 12
- 150000002653 sesterterpene derivatives Chemical group 0.000 claims description 11
- 239000013604 expression vector Substances 0.000 claims description 8
- 239000002773 nucleotide Substances 0.000 claims description 8
- 125000003729 nucleotide group Chemical group 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 238000006555 catalytic reaction Methods 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 238000001727 in vivo Methods 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 22
- 150000003505 terpenes Chemical class 0.000 abstract description 17
- 238000000034 method Methods 0.000 abstract description 15
- 102000004169 proteins and genes Human genes 0.000 abstract description 15
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 abstract description 9
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 abstract description 9
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 abstract description 8
- 230000035772 mutation Effects 0.000 abstract description 7
- 150000001413 amino acids Chemical group 0.000 abstract description 6
- 229930014626 natural product Natural products 0.000 abstract description 6
- 238000001514 detection method Methods 0.000 abstract description 3
- 238000010276 construction Methods 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 32
- 229940125782 compound 2 Drugs 0.000 description 23
- UHOVQNZJYSORNB-MICDWDOJSA-N deuteriobenzene Chemical compound [2H]C1=CC=CC=C1 UHOVQNZJYSORNB-MICDWDOJSA-N 0.000 description 23
- 229940126214 compound 3 Drugs 0.000 description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 238000010586 diagram Methods 0.000 description 18
- 238000005481 NMR spectroscopy Methods 0.000 description 17
- MGRJONRALKLFBS-YBSCOXHPSA-N preterpestacin I Chemical compound CC(C)[C@H](CC1)[C@@H](C/C=C2\C)[C@]1(C)C/C=C(\C)/CC/C=C(\C)/CC[C@@H]\2O MGRJONRALKLFBS-YBSCOXHPSA-N 0.000 description 14
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 101710135150 (+)-T-muurolol synthase ((2E,6E)-farnesyl diphosphate cyclizing) Proteins 0.000 description 8
- 101710119400 Geranylfarnesyl diphosphate synthase Proteins 0.000 description 8
- 101710107752 Geranylgeranyl diphosphate synthase Proteins 0.000 description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 8
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 8
- 108010050848 glycylleucine Proteins 0.000 description 8
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 description 8
- 235000007586 terpenes Nutrition 0.000 description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 7
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 7
- 230000014759 maintenance of location Effects 0.000 description 7
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 7
- SBHUBSDEZQFJHJ-CIUDSAMLSA-N Asp-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O SBHUBSDEZQFJHJ-CIUDSAMLSA-N 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 239000013612 plasmid Substances 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 5
- 241000371644 Curvularia ravenelii Species 0.000 description 5
- 241001052560 Thallis Species 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 238000005100 correlation spectroscopy Methods 0.000 description 5
- 239000013078 crystal Substances 0.000 description 5
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 5
- CBIDRCWHNCKSTO-UHFFFAOYSA-N prenyl diphosphate Chemical compound CC(C)=CCO[P@](O)(=O)OP(O)(O)=O CBIDRCWHNCKSTO-UHFFFAOYSA-N 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 238000004262 preparative liquid chromatography Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- NKBQZKVMKJJDLX-SRVKXCTJSA-N Arg-Glu-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NKBQZKVMKJJDLX-SRVKXCTJSA-N 0.000 description 4
- 241000190150 Bipolaris sorokiniana Species 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- KPNWAJMEMRCLAL-GUBZILKMSA-N Gln-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KPNWAJMEMRCLAL-GUBZILKMSA-N 0.000 description 4
- LRPXYSGPOBVBEH-IUCAKERBSA-N Glu-Gly-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O LRPXYSGPOBVBEH-IUCAKERBSA-N 0.000 description 4
- GNPVTZJUUBPZKW-WDSKDSINSA-N Gly-Gln-Ser Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O GNPVTZJUUBPZKW-WDSKDSINSA-N 0.000 description 4
- JQLFYZMEXFNRFS-DJFWLOJKSA-N Ile-Asp-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N JQLFYZMEXFNRFS-DJFWLOJKSA-N 0.000 description 4
- OHKFXGKHSJKKAL-NRPADANISA-N Ser-Glu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OHKFXGKHSJKKAL-NRPADANISA-N 0.000 description 4
- 108010068380 arginylarginine Proteins 0.000 description 4
- 125000002619 bicyclic group Chemical group 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 125000004122 cyclic group Chemical group 0.000 description 4
- 238000009510 drug design Methods 0.000 description 4
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 108010064235 lysylglycine Proteins 0.000 description 4
- 108010051242 phenylalanylserine Proteins 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 238000007363 ring formation reaction Methods 0.000 description 4
- 238000010898 silica gel chromatography Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 108010006731 Dimethylallyltranstransferase Proteins 0.000 description 3
- 102000005454 Dimethylallyltranstransferase Human genes 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 101100226889 Phomopsis amygdali PaFS gene Proteins 0.000 description 3
- 108010050516 adenylate isopentenyltransferase Proteins 0.000 description 3
- 230000002902 bimodal effect Effects 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N hexane Substances CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 238000010829 isocratic elution Methods 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 3
- 238000013508 migration Methods 0.000 description 3
- 230000005012 migration Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- YLTKNGYYPIWKHZ-ACZMJKKPSA-N Ala-Ala-Glu Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O YLTKNGYYPIWKHZ-ACZMJKKPSA-N 0.000 description 2
- NHCPCLJZRSIDHS-ZLUOBGJFSA-N Ala-Asp-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O NHCPCLJZRSIDHS-ZLUOBGJFSA-N 0.000 description 2
- CXZFXHGJJPVUJE-CIUDSAMLSA-N Ala-Cys-Leu Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)O)N CXZFXHGJJPVUJE-CIUDSAMLSA-N 0.000 description 2
- SFNFGFDRYJKZKN-XQXXSGGOSA-N Ala-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](C)N)O SFNFGFDRYJKZKN-XQXXSGGOSA-N 0.000 description 2
- BVSGPHDECMJBDE-HGNGGELXSA-N Ala-Glu-His Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N BVSGPHDECMJBDE-HGNGGELXSA-N 0.000 description 2
- OKIKVSXTXVVFDV-MMWGEVLESA-N Ala-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C)N OKIKVSXTXVVFDV-MMWGEVLESA-N 0.000 description 2
- VNYMOTCMNHJGTG-JBDRJPRFSA-N Ala-Ile-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O VNYMOTCMNHJGTG-JBDRJPRFSA-N 0.000 description 2
- CCDFBRZVTDDJNM-GUBZILKMSA-N Ala-Leu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O CCDFBRZVTDDJNM-GUBZILKMSA-N 0.000 description 2
- LDLSENBXQNDTPB-DCAQKATOSA-N Ala-Lys-Arg Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LDLSENBXQNDTPB-DCAQKATOSA-N 0.000 description 2
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 2
- GKAZXNDATBWNBI-DCAQKATOSA-N Ala-Met-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)O)N GKAZXNDATBWNBI-DCAQKATOSA-N 0.000 description 2
- BFMIRJBURUXDRG-DLOVCJGASA-N Ala-Phe-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 BFMIRJBURUXDRG-DLOVCJGASA-N 0.000 description 2
- XWFWAXPOLRTDFZ-FXQIFTODSA-N Ala-Pro-Ser Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O XWFWAXPOLRTDFZ-FXQIFTODSA-N 0.000 description 2
- VJVQKGYHIZPSNS-FXQIFTODSA-N Ala-Ser-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N VJVQKGYHIZPSNS-FXQIFTODSA-N 0.000 description 2
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 2
- KMSHNDWHPWXPEC-BQBZGAKWSA-N Arg-Asp-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O KMSHNDWHPWXPEC-BQBZGAKWSA-N 0.000 description 2
- OTCJMMRQBVDQRK-DCAQKATOSA-N Arg-Asp-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O OTCJMMRQBVDQRK-DCAQKATOSA-N 0.000 description 2
- TTXYKSADPSNOIF-IHRRRGAJSA-N Arg-Asp-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O TTXYKSADPSNOIF-IHRRRGAJSA-N 0.000 description 2
- YHQGEARSFILVHL-HJGDQZAQSA-N Arg-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N)O YHQGEARSFILVHL-HJGDQZAQSA-N 0.000 description 2
- MZRBYBIQTIKERR-GUBZILKMSA-N Arg-Glu-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MZRBYBIQTIKERR-GUBZILKMSA-N 0.000 description 2
- DJAIOAKQIOGULM-DCAQKATOSA-N Arg-Glu-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O DJAIOAKQIOGULM-DCAQKATOSA-N 0.000 description 2
- JAYIQMNQDMOBFY-KKUMJFAQSA-N Arg-Glu-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JAYIQMNQDMOBFY-KKUMJFAQSA-N 0.000 description 2
- QKSAZKCRVQYYGS-UWVGGRQHSA-N Arg-Gly-His Chemical compound N[C@@H](CCCN=C(N)N)C(=O)NCC(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O QKSAZKCRVQYYGS-UWVGGRQHSA-N 0.000 description 2
- ZZZWQALDSQQBEW-STQMWFEESA-N Arg-Gly-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZZZWQALDSQQBEW-STQMWFEESA-N 0.000 description 2
- LLUGJARLJCGLAR-CYDGBPFRSA-N Arg-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N LLUGJARLJCGLAR-CYDGBPFRSA-N 0.000 description 2
- OISWSORSLQOGFV-AVGNSLFASA-N Arg-Met-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCSC)NC(=O)[C@@H](N)CCCN=C(N)N OISWSORSLQOGFV-AVGNSLFASA-N 0.000 description 2
- MNBHKGYCLBUIBC-UFYCRDLUSA-N Arg-Phe-Phe Chemical compound C([C@H](NC(=O)[C@H](CCCNC(N)=N)N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MNBHKGYCLBUIBC-UFYCRDLUSA-N 0.000 description 2
- SLQQPJBDBVPVQV-JYJNAYRXSA-N Arg-Phe-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O SLQQPJBDBVPVQV-JYJNAYRXSA-N 0.000 description 2
- DDBMKOCQWNFDBH-RHYQMDGZSA-N Arg-Thr-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O DDBMKOCQWNFDBH-RHYQMDGZSA-N 0.000 description 2
- YNDLOUMBVDVALC-ZLUOBGJFSA-N Asn-Ala-Ala Chemical compound C[C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CC(=O)N)N YNDLOUMBVDVALC-ZLUOBGJFSA-N 0.000 description 2
- XWGJDUSDTRPQRK-ZLUOBGJFSA-N Asn-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(N)=O XWGJDUSDTRPQRK-ZLUOBGJFSA-N 0.000 description 2
- NTXNUXPCNRDMAF-WFBYXXMGSA-N Asn-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC(N)=O)C)C(O)=O)=CNC2=C1 NTXNUXPCNRDMAF-WFBYXXMGSA-N 0.000 description 2
- PNHQRQTVBRDIEF-CIUDSAMLSA-N Asn-Leu-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(=O)N)N PNHQRQTVBRDIEF-CIUDSAMLSA-N 0.000 description 2
- YUUIAUXBNOHFRJ-IHRRRGAJSA-N Asn-Phe-Met Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(O)=O YUUIAUXBNOHFRJ-IHRRRGAJSA-N 0.000 description 2
- XCBKBPRFACFFOO-AQZXSJQPSA-N Asn-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O XCBKBPRFACFFOO-AQZXSJQPSA-N 0.000 description 2
- MLJZMGIXXMTEPO-UBHSHLNASA-N Asn-Trp-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CO)C(O)=O MLJZMGIXXMTEPO-UBHSHLNASA-N 0.000 description 2
- KRXIWXCXOARFNT-ZLUOBGJFSA-N Asp-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O KRXIWXCXOARFNT-ZLUOBGJFSA-N 0.000 description 2
- VTYQAQFKMQTKQD-ACZMJKKPSA-N Asp-Ala-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O VTYQAQFKMQTKQD-ACZMJKKPSA-N 0.000 description 2
- GWTLRDMPMJCNMH-WHFBIAKZSA-N Asp-Asn-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GWTLRDMPMJCNMH-WHFBIAKZSA-N 0.000 description 2
- KNMRXHIAVXHCLW-ZLUOBGJFSA-N Asp-Asn-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N)C(=O)O KNMRXHIAVXHCLW-ZLUOBGJFSA-N 0.000 description 2
- WCFCYFDBMNFSPA-ACZMJKKPSA-N Asp-Asp-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCC(O)=O WCFCYFDBMNFSPA-ACZMJKKPSA-N 0.000 description 2
- VZNOVQKGJQJOCS-SRVKXCTJSA-N Asp-Asp-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VZNOVQKGJQJOCS-SRVKXCTJSA-N 0.000 description 2
- OMMIEVATLAGRCK-BYPYZUCNSA-N Asp-Gly-Gly Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)NCC(O)=O OMMIEVATLAGRCK-BYPYZUCNSA-N 0.000 description 2
- JXGJJQJHXHXJQF-CIUDSAMLSA-N Asp-Met-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O JXGJJQJHXHXJQF-CIUDSAMLSA-N 0.000 description 2
- ZBYLEBZCVKLPCY-FXQIFTODSA-N Asp-Ser-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O ZBYLEBZCVKLPCY-FXQIFTODSA-N 0.000 description 2
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 2
- SQIARYGNVQWOSB-BZSNNMDCSA-N Asp-Tyr-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQIARYGNVQWOSB-BZSNNMDCSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- AEJSNWMRPXAKCW-WHFBIAKZSA-N Cys-Ala-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O AEJSNWMRPXAKCW-WHFBIAKZSA-N 0.000 description 2
- ZVNFONSZVUBRAV-CIUDSAMLSA-N Cys-Gln-Arg Chemical compound C(C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CS)N)CN=C(N)N ZVNFONSZVUBRAV-CIUDSAMLSA-N 0.000 description 2
- VBPGTULCFGKGTF-ACZMJKKPSA-N Cys-Glu-Asp Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VBPGTULCFGKGTF-ACZMJKKPSA-N 0.000 description 2
- UCSXXFRXHGUXCQ-SRVKXCTJSA-N Cys-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CS)N UCSXXFRXHGUXCQ-SRVKXCTJSA-N 0.000 description 2
- DQUWSUWXPWGTQT-DCAQKATOSA-N Cys-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CS DQUWSUWXPWGTQT-DCAQKATOSA-N 0.000 description 2
- SWJYSDXMTPMBHO-FXQIFTODSA-N Cys-Pro-Ser Chemical compound [H]N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SWJYSDXMTPMBHO-FXQIFTODSA-N 0.000 description 2
- 238000003775 Density Functional Theory Methods 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- RZSLYUUFFVHFRQ-FXQIFTODSA-N Gln-Ala-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O RZSLYUUFFVHFRQ-FXQIFTODSA-N 0.000 description 2
- XOKGKOQWADCLFQ-GARJFASQSA-N Gln-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XOKGKOQWADCLFQ-GARJFASQSA-N 0.000 description 2
- DHNWZLGBTPUTQQ-QEJZJMRPSA-N Gln-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N DHNWZLGBTPUTQQ-QEJZJMRPSA-N 0.000 description 2
- OFPWCBGRYAOLMU-AVGNSLFASA-N Gln-Asp-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N)O OFPWCBGRYAOLMU-AVGNSLFASA-N 0.000 description 2
- APWLZZSLCXLDCF-CIUDSAMLSA-N Gln-Cys-Met Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCSC)C(O)=O APWLZZSLCXLDCF-CIUDSAMLSA-N 0.000 description 2
- MCAVASRGVBVPMX-FXQIFTODSA-N Gln-Glu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MCAVASRGVBVPMX-FXQIFTODSA-N 0.000 description 2
- TWTWUBHEWQPMQW-ZPFDUUQYSA-N Gln-Ile-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O TWTWUBHEWQPMQW-ZPFDUUQYSA-N 0.000 description 2
- VNTGPISAOMAXRK-CIUDSAMLSA-N Gln-Pro-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O VNTGPISAOMAXRK-CIUDSAMLSA-N 0.000 description 2
- OSCLNNWLKKIQJM-WDSKDSINSA-N Gln-Ser-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O OSCLNNWLKKIQJM-WDSKDSINSA-N 0.000 description 2
- SXFPZRRVWSUYII-KBIXCLLPSA-N Gln-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N SXFPZRRVWSUYII-KBIXCLLPSA-N 0.000 description 2
- LKDIBBOKUAASNP-FXQIFTODSA-N Glu-Ala-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LKDIBBOKUAASNP-FXQIFTODSA-N 0.000 description 2
- CVPXINNKRTZBMO-CIUDSAMLSA-N Glu-Arg-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)CN=C(N)N CVPXINNKRTZBMO-CIUDSAMLSA-N 0.000 description 2
- XKPOCESCRTVRPL-KBIXCLLPSA-N Glu-Cys-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XKPOCESCRTVRPL-KBIXCLLPSA-N 0.000 description 2
- CGOHAEBMDSEKFB-FXQIFTODSA-N Glu-Glu-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O CGOHAEBMDSEKFB-FXQIFTODSA-N 0.000 description 2
- LZMQSTPFYJLVJB-GUBZILKMSA-N Glu-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N LZMQSTPFYJLVJB-GUBZILKMSA-N 0.000 description 2
- JDUKCSSHWNIQQZ-IHRRRGAJSA-N Glu-Phe-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JDUKCSSHWNIQQZ-IHRRRGAJSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- DUYYPIRFTLOAJQ-YUMQZZPRSA-N Gly-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN DUYYPIRFTLOAJQ-YUMQZZPRSA-N 0.000 description 2
- SABZDFAAOJATBR-QWRGUYRKSA-N Gly-Cys-Phe Chemical compound [H]NCC(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SABZDFAAOJATBR-QWRGUYRKSA-N 0.000 description 2
- SWQALSGKVLYKDT-UHFFFAOYSA-N Gly-Ile-Ala Natural products NCC(=O)NC(C(C)CC)C(=O)NC(C)C(O)=O SWQALSGKVLYKDT-UHFFFAOYSA-N 0.000 description 2
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 2
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 2
- QGDOOCIPHSSADO-STQMWFEESA-N Gly-Met-Phe Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QGDOOCIPHSSADO-STQMWFEESA-N 0.000 description 2
- OMOZPGCHVWOXHN-BQBZGAKWSA-N Gly-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)CN OMOZPGCHVWOXHN-BQBZGAKWSA-N 0.000 description 2
- GJHWILMUOANXTG-WPRPVWTQSA-N Gly-Val-Arg Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GJHWILMUOANXTG-WPRPVWTQSA-N 0.000 description 2
- SYMSVYVUSPSAAO-IHRRRGAJSA-N His-Arg-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O SYMSVYVUSPSAAO-IHRRRGAJSA-N 0.000 description 2
- NOQPTNXSGNPJNS-YUMQZZPRSA-N His-Asn-Gly Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O NOQPTNXSGNPJNS-YUMQZZPRSA-N 0.000 description 2
- YADRBUZBKHHDAO-XPUUQOCRSA-N His-Gly-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](C)C(O)=O YADRBUZBKHHDAO-XPUUQOCRSA-N 0.000 description 2
- MPXGJGBXCRQQJE-MXAVVETBSA-N His-Ile-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O MPXGJGBXCRQQJE-MXAVVETBSA-N 0.000 description 2
- LVWIJITYHRZHBO-IXOXFDKPSA-N His-Leu-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LVWIJITYHRZHBO-IXOXFDKPSA-N 0.000 description 2
- BZAQOPHNBFOOJS-DCAQKATOSA-N His-Pro-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O BZAQOPHNBFOOJS-DCAQKATOSA-N 0.000 description 2
- PYNPBMCLAKTHJL-SRVKXCTJSA-N His-Pro-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O PYNPBMCLAKTHJL-SRVKXCTJSA-N 0.000 description 2
- JXUGDUWBMKIJDC-NAKRPEOUSA-N Ile-Ala-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JXUGDUWBMKIJDC-NAKRPEOUSA-N 0.000 description 2
- YKRIXHPEIZUDDY-GMOBBJLQSA-N Ile-Asn-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YKRIXHPEIZUDDY-GMOBBJLQSA-N 0.000 description 2
- WZDCVAWMBUNDDY-KBIXCLLPSA-N Ile-Glu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](C)C(=O)O)N WZDCVAWMBUNDDY-KBIXCLLPSA-N 0.000 description 2
- LPFBXFILACZHIB-LAEOZQHASA-N Ile-Gly-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)O)C(=O)O)N LPFBXFILACZHIB-LAEOZQHASA-N 0.000 description 2
- LNJLOZYNZFGJMM-DEQVHRJGSA-N Ile-His-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N2CCC[C@@H]2C(=O)O)N LNJLOZYNZFGJMM-DEQVHRJGSA-N 0.000 description 2
- NPAYJTAXWXJKLO-NAKRPEOUSA-N Ile-Met-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)O)N NPAYJTAXWXJKLO-NAKRPEOUSA-N 0.000 description 2
- UYNXBNHVWFNVIN-HJWJTTGWSA-N Ile-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)CC)CC1=CC=CC=C1 UYNXBNHVWFNVIN-HJWJTTGWSA-N 0.000 description 2
- PELCGFMHLZXWBQ-BJDJZHNGSA-N Ile-Ser-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)O)N PELCGFMHLZXWBQ-BJDJZHNGSA-N 0.000 description 2
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 2
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 2
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 2
- CZCSUZMIRKFFFA-CIUDSAMLSA-N Leu-Ala-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O CZCSUZMIRKFFFA-CIUDSAMLSA-N 0.000 description 2
- UILIPCLTHRPCRB-XUXIUFHCSA-N Leu-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(C)C)N UILIPCLTHRPCRB-XUXIUFHCSA-N 0.000 description 2
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 2
- MMEDVBWCMGRKKC-GARJFASQSA-N Leu-Asp-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N MMEDVBWCMGRKKC-GARJFASQSA-N 0.000 description 2
- PPBKJAQJAUHZKX-SRVKXCTJSA-N Leu-Cys-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC(C)C PPBKJAQJAUHZKX-SRVKXCTJSA-N 0.000 description 2
- YVKSMSDXKMSIRX-GUBZILKMSA-N Leu-Glu-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YVKSMSDXKMSIRX-GUBZILKMSA-N 0.000 description 2
- KVMULWOHPPMHHE-DCAQKATOSA-N Leu-Glu-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O KVMULWOHPPMHHE-DCAQKATOSA-N 0.000 description 2
- WIDZHJTYKYBLSR-DCAQKATOSA-N Leu-Glu-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WIDZHJTYKYBLSR-DCAQKATOSA-N 0.000 description 2
- NEEOBPIXKWSBRF-IUCAKERBSA-N Leu-Glu-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O NEEOBPIXKWSBRF-IUCAKERBSA-N 0.000 description 2
- SGIIOQQGLUUMDQ-IHRRRGAJSA-N Leu-His-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](C(C)C)C(=O)O)N SGIIOQQGLUUMDQ-IHRRRGAJSA-N 0.000 description 2
- ORWTWZXGDBYVCP-BJDJZHNGSA-N Leu-Ile-Cys Chemical compound SC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC(C)C ORWTWZXGDBYVCP-BJDJZHNGSA-N 0.000 description 2
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 2
- INCJJHQRZGQLFC-KBPBESRZSA-N Leu-Phe-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O INCJJHQRZGQLFC-KBPBESRZSA-N 0.000 description 2
- SYRTUBLKWNDSDK-DKIMLUQUSA-N Leu-Phe-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O SYRTUBLKWNDSDK-DKIMLUQUSA-N 0.000 description 2
- SQUFDMCWMFOEBA-KKUMJFAQSA-N Leu-Ser-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SQUFDMCWMFOEBA-KKUMJFAQSA-N 0.000 description 2
- JGKHAFUAPZCCDU-BZSNNMDCSA-N Leu-Tyr-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CC=C(O)C=C1 JGKHAFUAPZCCDU-BZSNNMDCSA-N 0.000 description 2
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 2
- VKVDRTGWLVZJOM-DCAQKATOSA-N Leu-Val-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O VKVDRTGWLVZJOM-DCAQKATOSA-N 0.000 description 2
- SJNZALDHDUYDBU-IHRRRGAJSA-N Lys-Arg-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(O)=O SJNZALDHDUYDBU-IHRRRGAJSA-N 0.000 description 2
- SSJBMGCZZXCGJJ-DCAQKATOSA-N Lys-Asp-Met Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O SSJBMGCZZXCGJJ-DCAQKATOSA-N 0.000 description 2
- GQFDWEDHOQRNLC-QWRGUYRKSA-N Lys-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN GQFDWEDHOQRNLC-QWRGUYRKSA-N 0.000 description 2
- RBEATVHTWHTHTJ-KKUMJFAQSA-N Lys-Leu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O RBEATVHTWHTHTJ-KKUMJFAQSA-N 0.000 description 2
- LUAJJLPHUXPQLH-KKUMJFAQSA-N Lys-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCCN)N LUAJJLPHUXPQLH-KKUMJFAQSA-N 0.000 description 2
- SBQDRNOLGSYHQA-YUMQZZPRSA-N Lys-Ser-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SBQDRNOLGSYHQA-YUMQZZPRSA-N 0.000 description 2
- JHNOXVASMSXSNB-WEDXCCLWSA-N Lys-Thr-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O JHNOXVASMSXSNB-WEDXCCLWSA-N 0.000 description 2
- DLCAXBGXGOVUCD-PPCPHDFISA-N Lys-Thr-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DLCAXBGXGOVUCD-PPCPHDFISA-N 0.000 description 2
- RMOKGALPSPOYKE-KATARQTJSA-N Lys-Thr-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O RMOKGALPSPOYKE-KATARQTJSA-N 0.000 description 2
- VVURYEVJJTXWNE-ULQDDVLXSA-N Lys-Tyr-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O VVURYEVJJTXWNE-ULQDDVLXSA-N 0.000 description 2
- UAPZLLPGGOOCRO-IHRRRGAJSA-N Met-Asn-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N UAPZLLPGGOOCRO-IHRRRGAJSA-N 0.000 description 2
- DJDFBVNNDAUPRW-GUBZILKMSA-N Met-Glu-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O DJDFBVNNDAUPRW-GUBZILKMSA-N 0.000 description 2
- GPAHWYRSHCKICP-GUBZILKMSA-N Met-Glu-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O GPAHWYRSHCKICP-GUBZILKMSA-N 0.000 description 2
- MVMNUCOHQGYYKB-PEDHHIEDSA-N Met-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H](CCSC)N MVMNUCOHQGYYKB-PEDHHIEDSA-N 0.000 description 2
- AFFKUNVPPLQUGA-DCAQKATOSA-N Met-Leu-Ala Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O AFFKUNVPPLQUGA-DCAQKATOSA-N 0.000 description 2
- PZUUMQPMHBJJKE-AVGNSLFASA-N Met-Leu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCNC(N)=N PZUUMQPMHBJJKE-AVGNSLFASA-N 0.000 description 2
- ZIIMORLEZLVRIP-SRVKXCTJSA-N Met-Leu-Gln Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZIIMORLEZLVRIP-SRVKXCTJSA-N 0.000 description 2
- USBFEVBHEQBWDD-AVGNSLFASA-N Met-Leu-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O USBFEVBHEQBWDD-AVGNSLFASA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- 108010079364 N-glycylalanine Proteins 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- VHWOBXIWBDWZHK-IHRRRGAJSA-N Phe-Arg-Asp Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 VHWOBXIWBDWZHK-IHRRRGAJSA-N 0.000 description 2
- SMFGCTXUBWEPKM-KBPBESRZSA-N Phe-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 SMFGCTXUBWEPKM-KBPBESRZSA-N 0.000 description 2
- IWZRODDWOSIXPZ-IRXDYDNUSA-N Phe-Phe-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(O)=O)C1=CC=CC=C1 IWZRODDWOSIXPZ-IRXDYDNUSA-N 0.000 description 2
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 2
- GLJZDMZJHFXJQG-BZSNNMDCSA-N Phe-Ser-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GLJZDMZJHFXJQG-BZSNNMDCSA-N 0.000 description 2
- APXXVISUHOLGEE-ILWGZMRPSA-N Phe-Trp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CC4=CC=CC=C4)N)C(=O)O APXXVISUHOLGEE-ILWGZMRPSA-N 0.000 description 2
- DBNGDEAQXGFGRA-ACRUOGEOSA-N Phe-Tyr-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DBNGDEAQXGFGRA-ACRUOGEOSA-N 0.000 description 2
- GLUYKHMBGKQBHE-JYJNAYRXSA-N Phe-Val-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 GLUYKHMBGKQBHE-JYJNAYRXSA-N 0.000 description 2
- HPXVFFIIGOAQRV-DCAQKATOSA-N Pro-Arg-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O HPXVFFIIGOAQRV-DCAQKATOSA-N 0.000 description 2
- HXOLCSYHGRNXJJ-IHRRRGAJSA-N Pro-Asp-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HXOLCSYHGRNXJJ-IHRRRGAJSA-N 0.000 description 2
- SOACYAXADBWDDT-CYDGBPFRSA-N Pro-Ile-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SOACYAXADBWDDT-CYDGBPFRSA-N 0.000 description 2
- LNOWDSPAYBWJOR-PEDHHIEDSA-N Pro-Ile-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LNOWDSPAYBWJOR-PEDHHIEDSA-N 0.000 description 2
- YXHYJEPDKSYPSQ-AVGNSLFASA-N Pro-Leu-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 YXHYJEPDKSYPSQ-AVGNSLFASA-N 0.000 description 2
- SWRNSCMUXRLHCR-ULQDDVLXSA-N Pro-Phe-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 SWRNSCMUXRLHCR-ULQDDVLXSA-N 0.000 description 2
- FNGOXVQBBCMFKV-CIUDSAMLSA-N Pro-Ser-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O FNGOXVQBBCMFKV-CIUDSAMLSA-N 0.000 description 2
- BGWKULMLUIUPKY-BQBZGAKWSA-N Pro-Ser-Gly Chemical compound OC(=O)CNC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 BGWKULMLUIUPKY-BQBZGAKWSA-N 0.000 description 2
- LNICFEXCAHIJOR-DCAQKATOSA-N Pro-Ser-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O LNICFEXCAHIJOR-DCAQKATOSA-N 0.000 description 2
- SXJOPONICMGFCR-DCAQKATOSA-N Pro-Ser-Lys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O SXJOPONICMGFCR-DCAQKATOSA-N 0.000 description 2
- KWMZPPWYBVZIER-XGEHTFHBSA-N Pro-Ser-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWMZPPWYBVZIER-XGEHTFHBSA-N 0.000 description 2
- FUOGXAQMNJMBFG-WPRPVWTQSA-N Pro-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 FUOGXAQMNJMBFG-WPRPVWTQSA-N 0.000 description 2
- 108010079005 RDV peptide Proteins 0.000 description 2
- 108010003201 RGH 0205 Proteins 0.000 description 2
- BKOKTRCZXRIQPX-ZLUOBGJFSA-N Ser-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N BKOKTRCZXRIQPX-ZLUOBGJFSA-N 0.000 description 2
- HBZBPFLJNDXRAY-FXQIFTODSA-N Ser-Ala-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O HBZBPFLJNDXRAY-FXQIFTODSA-N 0.000 description 2
- YUSRGTQIPCJNHQ-CIUDSAMLSA-N Ser-Arg-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O YUSRGTQIPCJNHQ-CIUDSAMLSA-N 0.000 description 2
- COAHUSQNSVFYBW-FXQIFTODSA-N Ser-Asn-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O COAHUSQNSVFYBW-FXQIFTODSA-N 0.000 description 2
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 2
- BYIROAKULFFTEK-CIUDSAMLSA-N Ser-Asp-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO BYIROAKULFFTEK-CIUDSAMLSA-N 0.000 description 2
- SMIDBHKWSYUBRZ-ACZMJKKPSA-N Ser-Glu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O SMIDBHKWSYUBRZ-ACZMJKKPSA-N 0.000 description 2
- UOLGINIHBRIECN-FXQIFTODSA-N Ser-Glu-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O UOLGINIHBRIECN-FXQIFTODSA-N 0.000 description 2
- DJACUBDEDBZKLQ-KBIXCLLPSA-N Ser-Ile-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O DJACUBDEDBZKLQ-KBIXCLLPSA-N 0.000 description 2
- NLOAIFSWUUFQFR-CIUDSAMLSA-N Ser-Leu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O NLOAIFSWUUFQFR-CIUDSAMLSA-N 0.000 description 2
- HEUVHBXOVZONPU-BJDJZHNGSA-N Ser-Leu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HEUVHBXOVZONPU-BJDJZHNGSA-N 0.000 description 2
- BYCVMHKULKRVPV-GUBZILKMSA-N Ser-Lys-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYCVMHKULKRVPV-GUBZILKMSA-N 0.000 description 2
- OLKICIBQRVSQMA-SRVKXCTJSA-N Ser-Ser-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OLKICIBQRVSQMA-SRVKXCTJSA-N 0.000 description 2
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 2
- QNBVFKZSSRYNFX-CUJWVEQBSA-N Ser-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CO)N)O QNBVFKZSSRYNFX-CUJWVEQBSA-N 0.000 description 2
- NADLKBTYNKUJEP-KATARQTJSA-N Ser-Thr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NADLKBTYNKUJEP-KATARQTJSA-N 0.000 description 2
- SDFUZKIAHWRUCS-QEJZJMRPSA-N Ser-Trp-Glu Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CO)N SDFUZKIAHWRUCS-QEJZJMRPSA-N 0.000 description 2
- DDPVJPIGACCMEH-XQXXSGGOSA-N Thr-Ala-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O DDPVJPIGACCMEH-XQXXSGGOSA-N 0.000 description 2
- LXWZOMSOUAMOIA-JIOCBJNQSA-N Thr-Asn-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O LXWZOMSOUAMOIA-JIOCBJNQSA-N 0.000 description 2
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 2
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 2
- XIULAFZYEKSGAJ-IXOXFDKPSA-N Thr-Leu-His Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 XIULAFZYEKSGAJ-IXOXFDKPSA-N 0.000 description 2
- MGJLBZFUXUGMML-VOAKCMCISA-N Thr-Lys-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MGJLBZFUXUGMML-VOAKCMCISA-N 0.000 description 2
- LKJCABTUFGTPPY-HJGDQZAQSA-N Thr-Pro-Gln Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O LKJCABTUFGTPPY-HJGDQZAQSA-N 0.000 description 2
- DOBIBIXIHJKVJF-XKBZYTNZSA-N Thr-Ser-Gln Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(N)=O DOBIBIXIHJKVJF-XKBZYTNZSA-N 0.000 description 2
- BBPCSGKKPJUYRB-UVOCVTCTSA-N Thr-Thr-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O BBPCSGKKPJUYRB-UVOCVTCTSA-N 0.000 description 2
- OGOYMQWIWHGTGH-KZVJFYERSA-N Thr-Val-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O OGOYMQWIWHGTGH-KZVJFYERSA-N 0.000 description 2
- GTNCSPKYWCJZAC-XIRDDKMYSA-N Trp-Asp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N GTNCSPKYWCJZAC-XIRDDKMYSA-N 0.000 description 2
- HLDFBNPSURDYEN-VHWLVUOQSA-N Trp-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N HLDFBNPSURDYEN-VHWLVUOQSA-N 0.000 description 2
- OWSRIUBVJOQHNY-IHPCNDPISA-N Trp-Lys-His Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC3=CN=CN3)C(=O)O)N OWSRIUBVJOQHNY-IHPCNDPISA-N 0.000 description 2
- IIJWXEUNETVJPV-IHRRRGAJSA-N Tyr-Arg-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N)O IIJWXEUNETVJPV-IHRRRGAJSA-N 0.000 description 2
- PRONOHBTMLNXCZ-BZSNNMDCSA-N Tyr-Leu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 PRONOHBTMLNXCZ-BZSNNMDCSA-N 0.000 description 2
- GITNQBVCEQBDQC-KKUMJFAQSA-N Tyr-Lys-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O GITNQBVCEQBDQC-KKUMJFAQSA-N 0.000 description 2
- QKXAEWMHAAVVGS-KKUMJFAQSA-N Tyr-Pro-Glu Chemical compound N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O QKXAEWMHAAVVGS-KKUMJFAQSA-N 0.000 description 2
- RGYCVIZZTUBSSG-JYJNAYRXSA-N Tyr-Pro-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O RGYCVIZZTUBSSG-JYJNAYRXSA-N 0.000 description 2
- DTWMJYGOUWNWEC-IHPCNDPISA-N Tyr-Trp-Cys Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CS)C(O)=O)C1=CC=C(O)C=C1 DTWMJYGOUWNWEC-IHPCNDPISA-N 0.000 description 2
- AEOFMCAKYIQQFY-YDHLFZDLSA-N Tyr-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AEOFMCAKYIQQFY-YDHLFZDLSA-N 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- ZLFHAAGHGQBQQN-GUBZILKMSA-N Val-Ala-Pro Natural products CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O ZLFHAAGHGQBQQN-GUBZILKMSA-N 0.000 description 2
- QPZMOUMNTGTEFR-ZKWXMUAHSA-N Val-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N QPZMOUMNTGTEFR-ZKWXMUAHSA-N 0.000 description 2
- BZMIYHIJVVJPCK-QSFUFRPTSA-N Val-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N BZMIYHIJVVJPCK-QSFUFRPTSA-N 0.000 description 2
- GVJUTBOZZBTBIG-AVGNSLFASA-N Val-Lys-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N GVJUTBOZZBTBIG-AVGNSLFASA-N 0.000 description 2
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 2
- ZRSZTKTVPNSUNA-IHRRRGAJSA-N Val-Lys-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)C(C)C)C(O)=O ZRSZTKTVPNSUNA-IHRRRGAJSA-N 0.000 description 2
- GBIUHAYJGWVNLN-AEJSXWLSSA-N Val-Ser-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N GBIUHAYJGWVNLN-AEJSXWLSSA-N 0.000 description 2
- ZLMFVXMJFIWIRE-FHWLQOOXSA-N Val-Trp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](C(C)C)N ZLMFVXMJFIWIRE-FHWLQOOXSA-N 0.000 description 2
- CFIBZQOLUDURST-IHRRRGAJSA-N Val-Tyr-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CS)C(=O)O)N CFIBZQOLUDURST-IHRRRGAJSA-N 0.000 description 2
- 108010081404 acein-2 Proteins 0.000 description 2
- 150000001336 alkenes Chemical class 0.000 description 2
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 2
- 108010008355 arginyl-glutamine Proteins 0.000 description 2
- 108010066119 arginyl-leucyl-aspartyl-serine Proteins 0.000 description 2
- 108010062796 arginyllysine Proteins 0.000 description 2
- 108010092854 aspartyllysine Proteins 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 238000005336 cracking Methods 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 108010060199 cysteinylproline Proteins 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 108010080575 glutamyl-aspartyl-alanine Proteins 0.000 description 2
- 108010057083 glutamyl-aspartyl-leucine Proteins 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- HPAIKDPJURGQLN-UHFFFAOYSA-N glycyl-L-histidyl-L-phenylalanine Natural products C=1C=CC=CC=1CC(C(O)=O)NC(=O)C(NC(=O)CN)CC1=CN=CN1 HPAIKDPJURGQLN-UHFFFAOYSA-N 0.000 description 2
- 108010075431 glycyl-alanyl-phenylalanine Proteins 0.000 description 2
- 108010050475 glycyl-leucyl-tyrosine Proteins 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 108010040030 histidinoalanine Proteins 0.000 description 2
- 108010036413 histidylglycine Proteins 0.000 description 2
- 108010092114 histidylphenylalanine Proteins 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 2
- 108010057821 leucylproline Proteins 0.000 description 2
- 108010054155 lysyllysine Proteins 0.000 description 2
- 108010068488 methionylphenylalanine Proteins 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 108010084525 phenylalanyl-phenylalanyl-glycine Proteins 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 102220197688 rs771978846 Human genes 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 108010048818 seryl-histidine Proteins 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 108010071207 serylmethionine Proteins 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 108010061238 threonyl-glycine Proteins 0.000 description 2
- 108010080629 tryptophan-leucine Proteins 0.000 description 2
- 108010020532 tyrosyl-proline Proteins 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 241000102059 Aporosa benthamiana Species 0.000 description 1
- 241001465318 Aspergillus terreus Species 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 238000004057 DFT-B3LYP calculation Methods 0.000 description 1
- 241000589989 Helicobacter Species 0.000 description 1
- 102220466681 Histone H4 transcription factor_S89G_mutation Human genes 0.000 description 1
- 108010025076 Holoenzymes Proteins 0.000 description 1
- VTAJIXDZFCRWBR-UHFFFAOYSA-N Licoricesaponin B2 Natural products C1C(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2)C(O)=O)C)(C)CC2)(C)C2C(C)(C)CC1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O VTAJIXDZFCRWBR-UHFFFAOYSA-N 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 101100186946 Neosartorya fischeri (strain ATCC 1020 / DSM 3700 / CBS 544.65 / FGSC A1164 / JCM 1740 / NRRL 181 / WB 181) NfSS gene Proteins 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 240000002044 Rhizophora apiculata Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 229930101531 artemisinin Natural products 0.000 description 1
- BLUAFEHZUWYNDE-NNWCWBAJSA-N artemisinin Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2OC(=O)[C@@H]4C BLUAFEHZUWYNDE-NNWCWBAJSA-N 0.000 description 1
- 229960004191 artemisinin Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- UHOVQNZJYSORNB-MZWXYZOWSA-N benzene-d6 Chemical compound [2H]C1=C([2H])C([2H])=C([2H])C([2H])=C1[2H] UHOVQNZJYSORNB-MZWXYZOWSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 125000002837 carbocyclic group Chemical group 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 239000007806 chemical reaction intermediate Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 150000002009 diols Chemical class 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 125000000567 diterpene group Chemical group 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- 239000001685 glycyrrhizic acid Substances 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002887 multiple sequence alignment Methods 0.000 description 1
- 238000000238 one-dimensional nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 102200006560 rs755322824 Human genes 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 229930003839 sesquiterpene dimer Natural products 0.000 description 1
- VQMHZILKNHUAMP-AISFCYSSSA-N sesterfisherol Chemical compound CC(C)[C@H]1CC[C@]2(C)C[C@@H]3[C@@H](C)CC[C@H]4[C@H](C)CC\C4=C(C)\[C@@]3(O)C[C@@H]12 VQMHZILKNHUAMP-AISFCYSSSA-N 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 description 1
- 125000003003 spiro group Chemical group 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P15/00—Preparation of compounds containing at least three condensed carbocyclic rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/007—Preparation of hydrocarbons or halogenated hydrocarbons containing one or more isoprene units, i.e. terpenes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/03—Carbon-oxygen lyases (4.2) acting on phosphates (4.2.3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
本发明涉及双功能萜合酶及其突变体以及催化产物5‑15环系二倍半萜类化合物。双功能萜合酶氨基酸序列如SEQ ID NO.1所示，双功能萜合酶突变体是将SEQ IDNO.1所示氨基酸序列的第89位置的丝氨酸(Serine)突变为亮氨酸(Leucine)后获得的如SEQ ID NO.3所示氨基酸序列对应的蛋白质。与现有技术相比，本发明通过双功能萜合酶高效精简突变库的构建和突变产物的检测，改造了双功能萜合酶的催化功能，获得了具丰富环系结构的萜类化合物，对有效扩充萜类天然产物库，增添更多具潜在药物价值萜类化合物后备军具有重要意义。
Description
技术领域
本发明属于生物工程技术领域，尤其是涉及一种双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物。
背景技术
萜合酶是萜类化合物生物合成途径中一类重要的酶，负责将萜类线性前体环化形成具有不同环系及对映立体中心的结构。萜合酶复杂精密的催化决定了萜类化合物丰富多样的结构骨架。迄今为止已存在超过80,000种萜类化合物，约占所有天然产物数量的1/3，具有广泛的生物学活性，其中不乏许多著名药物分子，如抗疟疾的青蒿素，抗肿瘤的紫杉醇，抗肝炎药物甘草酸等。
双功能萜合酶主要来源于真菌，同时具有异戊烯基转移酶(prenyltransferase，PT)结构域和萜类环化酶(terpene cyclase，TC)结构域，因此兼具萜类合成前体二甲基丙烯基焦磷酸(dimethylallyl diphosphate，DMAPP)和异戊烯基焦磷酸(isopentenyldiphosphate，IPP)首尾相连和环化的双重功能。目前为止，真菌来源双功能萜合酶的催化产物均为二萜和二倍半萜，且多为三环或四环。根据催化机制可知，PT结构域决定了催化合成萜类化合物的碳链长度，而TC结构域则对产物的环系骨架和立体构型起关键作用，且随着环化反应中间体上碳正离子的迁移，产物的环系结构也将逐渐趋于复杂。然而，由于高能碳正离子中间体的存在，环化过程瞬息万变、难以捕捉，也为双功能萜合酶催化机制的研究带来困难。同时，由于双功能萜合酶双结构域及多个柔性区域的存在，蛋白质晶体结构的获取和解析也十分困难，目前为止仍未有一例双功能萜合酶全酶晶体结构，即使是截短的TC结构域也仅有一例，因此，该领域中关于酶的结构和功能关系的了解较为有限。
蛋白质工程方法包括定向进化、理性设计和半理性设计。由于双功能萜合酶蛋白晶体的不足，目前仍缺乏对其空间结构和功能关系的深入认知，理性设计无从下手，且成功率较低，因此并不适用。定向进化是常用的对酶蛋白进行改造和筛选的有效手段，但其随机突变的特性往往导致突变库过于庞大，具有成本高、效率低、周期长等弊端。近年逐渐发展起来的半理性设计采用生物信息学的手段，通过同源蛋白序列比对、蛋白空间立体结构的比较，同时参考已知催化机制等信息，理性选择蛋白质的改造靶点和替代密码子，从而构建较为精简的突变体文库，更有针对性地改造蛋白质。因此，根据有限的双功能萜合酶结构和催化方面的信息，设计和构建高效突变体库，对更加深入了解萜合酶结构与功能的关系，有效扩充萜类天然产物库，增添更多具潜在药物价值萜类化合物后备军具有重要意义。
发明内容
本发明的目的在于提供一种双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物。
本发明通过对一种双功能萜合酶进行突变，得到双功能萜合酶突变体，该双功能萜合酶突变体能够催化复杂结构的二倍半萜类化合物。通过双功能萜合酶催化功能的改造和突变产物的获得，更加深入了解萜合酶结构与功能的关系，有效扩充萜类天然产物库，增添更多具潜在药物价值萜类化合物后备军。
本发明的目的可以通过以下技术方案来实现：
本发明首先提供一种双功能萜合酶，其氨基酸序列如SEQ ID NO.1所示。
本发明还提供一种双功能萜合酶突变体，是将SEQ ID NO.1所示氨基酸序列的第89位置的丝氨酸(Serine)突变为亮氨酸(Leucine)后获得的如SEQ ID NO.3所示氨基酸序列对应的蛋白质。
其中，SEQ ID NO.1所示氨基酸序列是筛选自小麦根腐平脐蠕孢菌Bipolarissorokiniana BS11134(保藏编号：CGMCC No.17767)的双功能萜合酶的氨基酸序列。其中，所述小麦根腐平脐蠕孢菌Bipolaris sorokiniana BS11134，已于2019年04月30日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC，地址：北京市朝阳区北辰西路1号院3号，中国科学院微生物研究所，邮编100101)，在中国专利CN110272345A中有记载。
本发明还提供一种分离的核酸，所述核酸编码所述双功能萜合酶，所述核酸的核苷酸序列如SEQ ID NO.2所示。
本发明还提供另一种分离的核酸，所述核酸编码所述双功能萜合酶突变体。该分离的核酸的核苷酸序列如SEQ ID NO.4所示。
本发明中提供的双功能萜合酶以及双功能萜合酶突变体是同时具有异戊烯基转移酶结构域和萜类环化酶结构域的二倍半萜合酶。
本发明还提供一种重组表达载体，所述重组表达载体包含编码双功能萜合酶或双功能萜合酶突变体的核酸。
本发明还提供一种重组表达转化体，所述重组表达转化体包含所述的重组表达载体。
本发明还提供双功能萜合酶突变体的催化产物，其中，双功能萜合酶突变体的催化产物2和3为含5-15-双环二倍半萜骨架的化合物，其分子式为C25H40，结构式分别如式2、式3所示；双功能萜合酶突变体的催化产物4，即化合物4为含5-6-8-5-四环二倍半萜骨架的化合物，双功能萜合酶突变体的催化产物5，即化合物5为含5-6-7-3-5-五环二倍半萜骨架的化合物，二者分子式均为C25H42O，结构式分别如式4、式5所示。
在本发明的一个实施方式中，双功能萜合酶突变体的催化产物是：双功能萜合酶突变体在酿酒酵母S.cerevisiae BJ5464中体内酶催化反应之后，提取的突变体催化产物。
由于采用上述技术方案，本发明具有以下优点和有益效果：
本发明提供了目标双功能萜合酶在酿酒酵母S.cerevisiae BJ5464中体内酶催化反应之后，提取、检测和制备突变体催化产物。
本发明提供了一种双功能萜合酶高效精简突变库的构建方法，以真菌来源第一例双功能萜合酶PaFS晶体结构为模板，构建BsPS蛋白结构模型，并通过POCASA预测活性催化口袋，排除掉高度保守位点和其他催化关键位点后，选定适当数量的突变位点，获得了“小而精”的突变体文库。
本发明通过双功能萜合酶高效精简突变库的构建和突变产物的检测，改造了双功能萜合酶的催化功能，获得了具丰富环系结构的萜类化合物，对有效扩充萜类天然产物库，增添更多具潜在药物价值萜类化合物后备军具有重要意义。
本发明的改造对象双功能萜合酶BsPS同时具有异戊烯基转移酶结构域以及萜类环化酶结构域，兼具萜类合成前体二甲基丙烯基焦磷酸(dimethylallyl diphosphate,DMAPP)和异戊烯基焦磷酸(isopentenyl diphosphate,IPP)首尾相连和环化的双重功能。BsPS的天然产物具有5-15双环的结构，该骨架结构可以通过碳正离子的迁移，形成其他同源蛋白催化复杂环产物。本发明通过改造BsPS，使突变蛋白能够催化5-6-8-5四环和5-6-7-3-5五环的复杂环二倍半萜化合物。
附图说明
图1为本发明实施例提供的双功能萜合酶蛋白模型的活性催化口袋及周围氨基酸残基示意图。
图2为本发明实施例提供的双功能萜合酶同源序列比对示意图。
图3为本发明实施例提供的双功能萜合酶原产物(1)和突变产物(化合物2-5)的化学结构图和GC-MS色谱示意图。
图4为本发明实施例提供的化合物2在甲醇中的HR-EI-MS示意图。
图5为本发明实施例提供的化合物2在Benzene-d6中的1H NMR示意图。
图6为本发明实施例提供的化合物2在Benzene-d6中的13C NMR示意图。
图7为本发明实施例提供的化合物2在Benzene-d6中的HSQC示意图。
图8为本发明实施例提供的化合物2在Benzene-d6中的1H-1H COSY示意图。
图9为本发明实施例提供的化合物2在Benzene-d6中的HMBC示意图。
图10为本发明实施例提供的化合物2在Benzene-d6中的NOSEY示意图。
图11为本发明实施例提供的化合物2的相对构型以及关键二维NMR相关。
图12为本发明实施例提供的化合物3在甲醇中的HR-EI-MS示意图。
图13为本发明实施例提供的化合物3在Benzene-d6中的1H NMR示意图。
图14为本发明实施例提供的化合物3在Benzene-d6中的13C NMR示意图。
图15为本发明实施例提供的化合物3在Benzene-d6中的HSQC示意图。
图16为本发明实施例提供的化合物3在Benzene-d6中的1H-1H COSY示意图。
图17为本发明实施例提供的化合物3在Benzene-d6中的HMBC示意图。
图18为本发明实施例提供的化合物3在Benzene-d6中的NOSEY示意图。
图19为本发明实施例提供的化合物3的相对构型以及关键二维NMR相关。
图20为本发明实施例提供的化合物4在甲醇中的HR-EI-MS示意图。
图21为本发明实施例提供的化合物4在Benzene-d6中的1H NMR示意图。
图22为本发明实施例提供的化合物4在Benzene-d6中的13C NMR示意图。
图23为本发明实施例提供的化合物4在Benzene-d6中的HSQC示意图。
图24为本发明实施例提供的化合物4在Benzene-d6中的1H-1H COSY示意图。
图25为本发明实施例提供的化合物4在Benzene-d6中的HMBC示意图。
图26为本发明实施例提供的化合物4在Benzene-d6中的NOSEY示意图。
图27为本发明实施例提供的化合物4的相对构型以及关键二维NMR相关。
图28为本发明实施例提供的化合物5在甲醇中的HR-EI-MS示意图。
图29为本发明实施例提供的化合物5在Benzene-d6中的1H NMR示意图。
图30为本发明实施例提供的化合物5在Benzene-d6中的13C NMR示意图。
图31为本发明实施例提供的化合物5在Benzene-d6中的HSQC示意图。
图32为本发明实施例提供的化合物5在Benzene-d6中的1H-1H COSY示意图。
图33为本发明实施例提供的化合物5在Benzene-d6中的HMBC示意图。
图34为本发明实施例提供的化合物5在Benzene-d6中的NOSEY示意图。
图35为本发明实施例提供的化合物5的相对构型以及关键二维NMR相关。
图36为本发明实施例提供的化合物2-5的关键NOE相关示意图。
图37为本发明实施例提供的化合物4的实际测定CD及计算ECD示意图。
图38为种双功能萜合酶的第89位丝氨酸突变为亮氨酸后，催化产物结构种类从单一的5-15双环，变为包括5-15双环、5-6-8-6四元环和5-6-8-3-5五元环等结构的二倍半萜化合物的示意图。
具体实施方式
为了更清楚地说明本发明，下面结合优选实施例对本发明做进一步的说明。本领域技术人员应当理解，下面所具体描述的内容是说明性的而非限制性的，不应以此限制本发明的保护范围。
实施例1
本实施例首先提供一种双功能萜合酶，其氨基酸序列如SEQ ID NO.1所示。
本实施例还提供一种双功能萜合酶突变体，是将SEQ ID NO.1所示氨基酸序列的第89位置的丝氨酸(Serine)突变为亮氨酸(Leucine)后获得的如SEQ ID NO.3所示氨基酸序列对应的蛋白质。
其中，SEQ ID NO.1所示氨基酸序列是筛选自小麦根腐平脐蠕孢菌Bipolarissorokiniana BS11134(保藏编号：CGMCC No.17767)的双功能萜合酶的氨基酸序列。其中，所述小麦根腐平脐蠕孢菌Bipolaris sorokiniana BS11134，已于2019年04月30日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC，地址：北京市朝阳区北辰西路1号院3号，中国科学院微生物研究所，邮编100101)，在中国专利CN110272345A中有记载。
本实施例提供一种分离的核酸，所述核酸编码本实施例中双功能萜合酶，所述双功能萜合酶的核苷酸序列如SEQ ID NO.2所示。该核酸的核苷酸序列是通过提取小麦根腐平脐蠕孢菌Bipolaris sorokiniana BS11134(保藏编号：CGMCC No.17767)的RNA，通过反转录的方法获得cDNA，然后测序得到。
本实施例还提供另一种分离的核酸，所述核酸编码本实施例中双功能萜合酶突变体。该分离的核酸的核苷酸序列如SEQ ID NO.4所示。其中，序列SEQ ID NO.4是通过PCR的方法将SEQ ID NO.2的265-267位碱基从TCT突变为CTT得到。
本实施例中提供的双功能萜合酶以及双功能萜合酶突变体是同时具有异戊烯基转移酶结构域和萜类环化酶结构域的二倍半萜合酶。
本实施例继续提供双功能萜合酶突变体的制备方法，包括以下步骤：
从小麦根腐平脐蠕孢菌Bipolaris sorokiniana BS11134(保藏编号：CGMCCNo.17767)的双功能萜合酶基因中调取双功能萜合酶BsPS编码基因的核苷酸序列和蛋白质序列，导入BLAST(https://blast.ncbi.nlm.nih.gov/Blast.cgi)中，并在PDB(http://www.rcsb.org/)数据库中进行检索，得到相似度为40％的真菌来源首例双功能萜合酶PaFS晶体结构，以此作为模板，应用SWISS-MODEL(https://swissmodel.expasy.org/)在线服务器，对双功能萜合酶BsPS的TC结构域进行蛋白模型的构建。再通过POCASA预测BsPS蛋白模型的活性催化口袋。
使用Clustal Omega(https://www.ebi.ac.uk/Tools/msa/clustalo/)对BsPS与其他已报道的双功能萜合酶PaFS、NfSS进行多序列比对。根据结构-序列比对结果，BsPS的第89位氨基酸位于催化口袋附近(图1)。亮氨酸在双功能萜合酶同源蛋白的催化口袋内出现频率高(图2)，因此通过PCR的方法将BsPS的第89位置的丝氨酸突变为亮氨酸(本领域技术人员均能通过PCR方法实现BsPS的第89位置的丝氨酸突变为亮氨酸)。
将双功能萜合酶的编码基因与线性化质粒pET28a片段重组，再将重组产物转化至大肠杆菌E.coli BL21(DE3)。之后将突变位点S89L设计在引物中进行全质粒PCR，使用DpnI对PCR产物消化以除去质粒模板，取2μL消化产物转化大肠杆菌E.coli BL21(DE3)，抗性平板筛选后，挑取单克隆并测序验证。获得突变体的pET28a重组质粒后，再将突变体编码基因与线性化质粒pYET-xw55片段重组，获得载有双功能萜合酶BsPS编码基因的重组质粒pYET-XW55-BsPS，再将重组产物转化至酿酒酵母S.cerevisiae BJ5464，挑取单克隆并测序验证，正确的菌株将用来后续发酵培养。
至此，本实施例提供了一种重组表达载体，所述重组表达载体包含编码本实施例中双功能萜合酶突变体的核酸。
至此，本实施例提供了一种重组表达转化体，所述重组表达转化体包含所述的重组表达载体。
本实施例还提供一种双功能萜合酶催化产物二倍半萜类化合物，包括：
双功能萜合酶的催化产物1为含5-15双环二倍半萜骨架的化合物，其分子式为C25H40，结构式如式(1)所示，命名为：Preterpestacin I。
本实施例双功能萜合酶突变体的催化产物2和3为含5-15-双环二倍半萜骨架的化合物，其分子式为C25H40，结构式分别如式2、式3所示；所述双功能萜合酶突变体的催化产物4，即化合物4为含5-6-8-5-四环二倍半萜骨架的化合物，所述双功能萜合酶突变体的催化产物5，即化合物5为含5-6-7-3-5-五环二倍半萜骨架的化合物，二者分子式均为C25H42O，结构式分别如式4、式5所示。
双功能萜合酶与双功能萜合酶突变体在酿酒酵母Saccharomyces cerevisiaeBJ5464中异源表达。
本实施例提供一种双功能萜合酶催化产物二倍半萜类化合物的制备方法包括以下步骤：
宿主菌为酿酒酵母S.cerevisiae BJ5464：以含双功能萜合酶编码基因工程菌经发酵培养获得菌体体内的双功能萜合酶为催化剂，直接从菌体内提取萜类产物，并通过GC-MS检测。
湿菌体为含双功能萜合酶编码基因的重组酿酒酵母S.cerevisiae BJ5464，发酵培养方法为含双功能萜合酶编码基因的重组酿酒酵母S.cerevisiae BJ5464接种于尿嘧啶缺陷型培养基(5g/L Bacto technical grade casamino acids，20g/L glucose，6.7g/LDifco yeast nitrogen base without amino acid,0.02g/L Tryptophan,0.02g/LAdenine)中，30℃、200rpm震荡培养48h，以2％的接种量接种于YPD培养基(10g/L yeastextract,20g/L peptone,20g/L glucose)，于30℃、200rpm震荡培养72h，获得酿酒酵母发酵湿菌体。
丙酮600mL超声(130W,30min)裂解酿酒酵母发酵湿菌体(1,100g)30min，离心后裂解上清，使用乙酸乙酯300mL涡旋振荡萃取3次，获得发酵培养的菌体粗提物1.5g。
发酵培养的菌体粗提物使用硅胶柱层析和半制备型液相色谱(HPLC)纯化和制备，包括以下步骤：
发酵培养的菌体粗提物1.5g使用硅胶柱层析进行样品初步分离。正己烷作为硅胶柱层析的洗脱流动相，通过TLC监测，获得含有化合物2和3的组分90mg。然后换用1：1的正己烷和二氯甲烷作为硅胶柱层析的洗脱流动相，通过TLC监测，分别获得含有化合物4和5的组分160mg和130mg。
硅胶柱洗脱下来含有目标化合物2、3、4、5的组分再经过半制备液相色谱柱进行分离纯化。含有化合物2和化合物3的组分，使用半制备液相色谱柱ACE Excel 5C18(Φ10×250mm)，100％乙腈，流速4mL/min等度洗脱制备，化合物2的保留时间为44min，化合物3的保留时间为43min。含有化合物4的组分，先使用半制备液相色谱柱ACE Excel 5C18-AR(Φ10×250mm)，100％乙腈，流速4mL/min等度洗脱，化合物4的保留时间为16.5min。接着，使用色谱柱COSMOSIL Cholester(Φ4.6×250mm)，100％甲醇，流速0.8mL/min等度洗脱纯化。含有化合物5的组分，先使用半制备液相色谱柱ACE Excel 5C18(Φ10×250mm)，100％乙腈，流速4mL/min等度洗脱，化合物5的保留时间为9min。接着，使用色谱柱COSMOSIL Cholester(Φ4.6×250mm)，100％甲醇，流速0.8mL/min等度洗脱纯化。
制备得到的化合物2～5使用气相色谱-质谱联用(GC-MS)检测，检测条件为：高压分流模式进样，进样口压力为110kPa，分流比为50，柱流量为1.77mL/min。起始温度60℃，之后以15℃/min的速率升至280℃，再以5℃/min的速率升至310℃，并在310℃保持13min。化合物2的保留时间为15.515min，m/z 340；化合物3的保留时间为15.625min，m/z 340；化合物4的保留时间为16.811min，m/z 358；化合物5的保留时间为16.592min，m/z 358。
双功能萜合酶BsPS突变催化产物和GC-MS色谱图如图3所示。与野生型相比，BsPS-S89A/S89C/S89G/S89L的突变产物中均获得了含5-15-双环二倍半萜骨架的化合物2和3，其分子式为C25H40；BsPS-S89L中获得了骨架结构更为丰富的化合物4和5，化合物4为含5-6-8-5-四环二倍半萜骨架的化合物，所述化合物5为含5-6-7-3-5-五环二倍半萜骨架的化合物，二者分子式均为C25H42O。
化合物2为白色粉状，HR-EI-MS测得其准分子离子峰为m/z 340.3125[M+](计算值为340.3130，误差值为1.5ppm)，不饱和度为6，分子式为C25H40(图4)。化合物2的1H NMR、13CNMR和HSQC NMR谱图数据分析显示：该化合物具有5个烯烃次甲基(δC/H 123.4/5.39；126.8/5.06；129.8/5.34；137.7/6.01；126.9/5.48)、3个烯烃季碳(δC 134.4；132.3；134.2)、1个sp3季碳(δC 47.3)、3个次甲基(δC/H 53.5/2.61；52.6/1.62；29.6/1.62)、7个亚甲基(δC/H44.6/2.36&1.94；40.9/2.27&2.02；24.7/2.29&2.03；40.3/2.02&1.97；25.1/2.22&2.06；43.5/1.48&1.35；30.2/1.37)、四个单峰甲基(δC/H 15.6/1.56；15.3/1.53；12.6/1.72；23.4/0.83)和两个双峰甲基(δC/H 22.9/1.01；22.4/0.94)(图5-7)。这些谱图特征与文献已报道的preterpestacin I(1)具有高度相似性，但化合物2多出一个双键，因此可以推测化合物2为二倍半萜化合物。
1H-1H COSY谱图表明该化合物具有4个自旋体系：H-1(δH 2.36&1.94)和H-2(δH5.39)构成自旋体系A；H-4(δH 2.27&2.02)、H-5(δH 2.29&2.03)和H-6(δH 5.06)构成自旋体系B；H-8(δH 2.02&1.97)、H-9(δH 2.22&2.06)和H-10(δH 5.34)构成自旋体系C；H-12(δH6.01)、H-13(δH 5.48)、H-14(δH 2.61)、H-18(δH 1.62)、H-17(δH 1.37)和H-16(δH 1.48&1.35)，以及通过H-19(δH 1.62)和H-18相连接的异丙基单元H-24(δH 1.01)和H-25(δH0.94)构成自旋体系D(图8)。HMBC谱图显示：H-20(δH 1.56)到C-2(δC 123.4)、C-3(δC134.4)和C-4(δC 40.9)之间的相关信号表明自旋体系A和B之间通过C-3相连接，H-21(δH1.53)到C-6(δC 126.8)、C-7(δC 132.3)和C-8(δC 40.3)之间的相关信号表明自旋体系B和C之间通过C-7相连接，H-22(δH 1.72)到C-10(δC 129.8)、C-11(δC 134.2)和C-12(δC 137.7)之间的相关信号表明自旋体系C和D之间通过C-11相连接，H-23(δH 0.83)到C-1(δC 44.6)、C-14(δC 53.5)、C-15(δC 47.3)和C-16(δC 43.5)之间的相关信号表明自旋体系D和A之间通过C-15相连接(图9)。综合上述谱图数据可知，化合物2为5-15双环结构的二倍半萜化合物。
NOESY谱图中的相关信号表明：化合物2中双键C2-C3、C6-C7和C10-C11均为E构型，双键C12-C13为反式构型(3JH12-H13＝15.0)。另外，H-23和H-25、H-13之间，H-1和H-14之间的NOE效应表明：23-CH3和与C-18相连接的异丙基单元(C24-C19-C25)为同一朝向，而H-14和H-1为相反朝向(图10，图36)。因此，化合物2的绝对构型可确定为14R,15S,18S-2(图11)。
化合物3同样为白色粉末，经HR-EI-MS测定其准分子离子峰为m/z 340.3126[M+](计算值为340.3130，误差值为1.2ppm)，不饱和度为6，分子式为C25H40(图12)。化合物3的1HNMR和13C NMR谱图特征与化合物2具有高度相似性，但化合物3多出一个末端双键取代了化合物2中的sp2次甲基(图13-14)。HMBC谱图中，H-20(δH 4.91)到C-2(δC 30.8)、C-3(δC150.4)和C-4(δC 37.7)之间，H-6(δH 5.08)到C-4之间，H-1(δH 1.54/1.47)到C-2之间，以及H-23到C-1、C-14(δC 56.3)、C-15(δC 46.3)和C-16(δC 40.9)之间的相关信号表明(图17)：化合物3是化合物2的位置异构体，在C20-C3位置存在一个末端双键。化合物3的NOESY谱图中信号相关(图18，图36)与化合物2高度相似，因此化合物3中的双键C6-C7、C10-C11和C12-C13均为E构型。由于化合物3相比于化合物2仅存在C2-C3位置双键的迁移，且C14-C18的化学位移相近，因此可确定二者的手性中心构型一致。所以化合物3的绝对构型可确定为14R,15S,18S-3(图19)。
化合物4为白色粉末，经HR-EI-MS测定其准分子离子峰为m/z 358.3240[M+](计算值为358.3236，误差值为1.1ppm)，不饱和度为5，分子式为C25H42O(图20)。化合物4的1H NMR、13C NMR和HSQC NMR谱图数据分析显示：该化合物具有2个烯烃季碳(δC 144.1；133.3)、1个氧取代季碳(δC 75.2)、1个sp3季碳(δC 42.0)、7个次甲基(δC/H 49.8/2.02；46.8/2.53；39.2/1.92；47.9/2.40；51.3/1.41；47.6/1.65；31.8/1.54)、8个亚甲基(δC/H 43.6/1.81&0.80；36.1/1.73&1.58；25.1/1.56；31.7/1.59；30.1/2.18&2.06；29.8/1.62；41.6/1.50&1.11；28.6/1.85&1.57)、3个单峰甲基(δC/H 32.6/1.28；15.0/1.68；19.4/0.80)和3个双峰甲基(δC/H 15.7/0.95；24.3/0.93；22.8/0.87)(图21-23)。这些谱图特征与NfSS催化合成的5-6-8-5四环结构二倍半萜sesterfisherol较为接近，因此化合物4可能也具有相同的碳环结构。HMBC谱图显示H-20(δH 1.28)到C-2(δC 49.8)、C-3(δC 75.2)和C-4(δC 36.1)之间，以及H-22(δH1.68)到C-10(δC 144.1)、C-11(δC 133.3)和C-12(δC 47.9)之间具有相关信号(图25)，因此，可以推定化合物4结构中的羟基基团连接在C-4位置上，是sesterfisherol的位置异构体。
NOESY谱图中H-25和H-23、H-24和H-13、H-12和H-14、H-12和H-6、H-21和H-5、H-20和H-1以及H-2和H-23之间存在NOE效应(图26，图36)，表明这些质子之间具有相同朝向，由此可以确定化合物4的相对构型。为进一步确定绝对构型，接下来使用TD-DFT(time-dependent density functional theory)方法进行ECD计算和模拟，计算方法和采用基组为B3LYP/6-31+G(d)。使用Sybyl 2.0软件进行初步构象分布搜索，使用Gaussian 09和DFT进行最小几何优化。最终，将化合物4的CD测试结果与ECD计算结果比对后(图37)，确定其绝对构型为2R,3R,6R,7R,12R,14R,15S,18S-4(图27)。
化合物5为白色粉末，经HR-EI-MS测定其准分子离子峰为m/z 358.3234[M+](计算值为358.3236，误差值为0.5ppm)，分子式为C25H42O(图28)。化合物5的1H NMR、13C NMR和HSQC NMR谱图数据分析显示：该化合物具有1个氧取代季碳(δC 75.3)、3个sp3季碳(δC41.3；30.8；41.2)、7个次甲基(δC/H 44.6/1.86；46.0/2.20；38.7/0.65；48.1/0.62；50.2/1.22；47.5/1.71；31.2/1.63)、8个亚甲基(δC/H 42.2/1.98&0.91；44.2/1.69&1.51；29.8/1.79&1.60；34.0/1.73；25.4/1.88&1.67；27.2/1.74&1.66；41.9/1.54&1.10；28.3/1.82&1.54)、3个单峰甲基(δC/H 24.3/1.06；11.9/1.16；19.2/0.83)和3个双峰甲基(δC/H 16.2/1.15；24.6/1.02；22.4/0.89)(图29-31)。这些谱图特征与红树林内生真菌Aspergillusterreus H010所产生的二倍半萜类化合物Aspterpenacid B较为接近，因此化合物5可能也具有5-3-7-6-5五环结构。但化合物5的1D NMR中未能观察到类似于Aspterpenacid B中羧基和氧取代次甲基的共振信号，而是多出一个甲基单峰和sp3次甲基。HMBC谱图显示H-22(δH1.16)到C-6(δC 41.3)、C-10(δC 38.7)、C-11(δC 30.8)和C-12(δC 48.1)之间，H-20(δH1.06)到C-2(δC 44.6)和C-3(δC75.3)之间，以及H-2(δH 1.86)到C-11和C-3之间具有相关信号(图33)，因此可以推定化合物5具有和Aspterpenacid B一致的碳环骨架，但其C-11位上为甲基基团，C-3位被羟基取代。化合物5的NOESY谱图数据显示，H-7和H-5、H-5和H-10、H-5和H-12、H-22和H-23、H-20和H-12、H-2和H-23、H-23和H-19、H-23和H-13以及H-13和H-24之间存在NOE效应(图34，图36)，再结合化合物1的2D NMR数据可知，化合物5和化合物1在C14、C15和C18位具有相同的立体化学特征，因此可推定化合物5的绝对构型为2R,3R,6R,7R,10R,11R,12R,14R,15S,18S-5(图35)。
上述的对实施例的描述是为便于该技术领域的普通技术人员能理解和使用发明。熟悉本领域技术的人员显然可以容易地对这些实施例做出各种修改，并把在此说明的一般原理应用到其他实施例中而不必经过创造性的劳动。因此，本发明不限于上述实施例，本领域技术人员根据本发明的揭示，不脱离本发明范畴所做出的改进和修改都应该在本发明的保护范围之内。
序列表
<110> 华东理工大学
<120> 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 702
<212> PRT
<213> Bipolaris sorokiniana
<400> 1
Met Glu Gln Leu Ser Tyr Gln Ser Lys Leu Ile Cys Ser Glu Glu Ser
1 5 10 15
Arg Gln Thr Gly Cys Phe Thr Thr Leu Pro Ile Arg Ile His Pro Arg
20 25 30
Asp Asp Leu Ala Asp Ala Ala Ser Arg Arg Phe Val Gln Asp Trp Ala
35 40 45
Arg Glu Met Arg Asp Gly Arg Glu Gln Ser Thr His Phe Ser Phe Ser
50 55 60
Pro Val Gly Asn Trp Ser Ser Leu Ile Tyr Pro Glu Ala Ile Pro Glu
65 70 75 80
Arg Leu Gly Val Leu Ala Tyr Leu Ser Asp Leu Gly Leu Ile His Asp
85 90 95
Asp Gly Gly Glu Gly Leu Ser Ile Glu Asp Ala Gln Ala Glu His Gly
100 105 110
Glu Leu Cys Ala Ala Leu Asp Pro Ser Asp Ile Ser Ser Val Ala Pro
115 120 125
Asp Ser Arg Ala Met Lys Thr Lys Lys Leu Val Ser Gln Cys Met Leu
130 135 140
Glu Cys Ile Ser Leu Asp Arg Glu Leu Gly Leu Lys Met Leu Ala Ala
145 150 155 160
Phe Arg Asp Val Trp Leu Ala Ile Ser Glu Arg Asn Ser Asp Lys Glu
165 170 175
Ala Gln Thr Met Glu Glu Tyr Leu Lys Tyr Arg Ser Asp Asn Gly Gly
180 185 190
Met Leu Val Phe Trp Pro Met Leu Gln Phe Ser Leu Gly Met Ser Ile
195 200 205
Ser Glu Ala Glu Glu Ala Leu Val Gln Pro Ile Ile Asp Ala Ala Thr
210 215 220
Glu Gly Leu Leu Leu Ala Asn Asp Tyr Phe Ser Trp Glu Arg Glu Tyr
225 230 235 240
Arg Glu Leu Gln Ser Gly Gln Ser Lys Arg Ile Val Ser Ala Val Asp
245 250 255
Leu Phe Ile Arg Thr Lys Gly Leu Ser Ile Asp His Ala Lys Glu Glu
260 265 270
Val Lys Arg Met Ile Ile Ala Ala Glu Arg Asp Phe Cys Gln Arg Arg
275 280 285
Asp Asp Leu Tyr Lys Asn His Pro Asp Ile Ser Leu Lys Leu Lys Arg
290 295 300
Trp Ile Asp Cys Ala Gly Leu Ala Val Ser Gly Asn His Tyr Trp Cys
305 310 315 320
Ser Ala Cys Pro Arg Gln Asn Ala Trp Lys Asp Met Thr Ser Gln Ser
325 330 335
His Asn Gly Ala Lys Arg Lys Thr Ser His Gly Ala Thr Ile Ser Met
340 345 350
Gln Glu Ala Pro Phe Lys Lys Arg Lys Asp Ser Ser Phe Phe Gly Ser
355 360 365
Gln Pro Ser Asp Asp Glu Pro Ser Leu Ser Glu Val Ser Ser Tyr Pro
370 375 380
Phe Tyr Lys Pro Ser Gly Leu Ala Leu Glu Ala Pro Ser Lys Tyr Val
385 390 395 400
Ser Asn Met Pro Ser Lys Gly Val Arg Ser Thr Leu Ile Glu Ala Leu
405 410 415
Asn Thr Trp Leu His Val Pro Ser Glu Arg Leu Asp Ser Ile Met Ser
420 425 430
Val Ile Asn Thr Leu His Asn Ala Ser Leu Ile Leu Asp Asp Leu Glu
435 440 445
Asp Asn Ser Pro Leu Arg Arg Gly Tyr Pro Ser Thr His Ile Leu Phe
450 455 460
Gly Gln Ser Gln Ser Ile Asn Ala Ala Asn Phe Met Phe Val Arg Ala
465 470 475 480
Val Gln Glu Val Ala Gln Asn Leu Ser Pro Asn Ala Leu Val Ala Val
485 490 495
Leu Glu Glu Leu Glu Gly Leu Tyr Leu Gly Gln Ser Trp Asp Leu Tyr
500 505 510
Trp Lys His Asn Leu Ala Cys Pro Ser Glu Ala Glu Tyr Val Asn Met
515 520 525
Ile Asp His Lys Thr Gly Gly Met Phe Arg Met Leu Leu Arg Ile Met
530 535 540
Gln Ala Glu Ser Glu Val Thr Pro Gln Pro Asp Phe His Arg Leu Thr
545 550 555 560
Leu Leu Phe Gly Arg Phe Phe Gln Ile Arg Asp Asp Tyr Met Asn Phe
565 570 575
Gln Asp Tyr Thr Ala Gln Lys Gly Leu Cys Glu Asp Leu Asp Glu Gly
580 585 590
Lys Phe Ser Tyr Pro Val Val Tyr Cys Leu Glu Asn His Pro Glu Tyr
595 600 605
Arg Gly His Phe Leu Gly Ile Phe Arg Gln Arg Pro Thr Val Ala Thr
610 615 620
Val Asn Ala Cys Pro Leu Ser Arg Glu Ser Lys Gln His Leu Thr Ala
625 630 635 640
Cys Leu Lys Lys Ser Gly Ala Phe Asp Lys Thr Ile Ala Cys Leu Met
645 650 655
Asp Met Glu Arg Asp Leu Glu Phe Glu Ile Asn Arg Leu Glu Gln Gln
660 665 670
Thr Gly Glu Thr Asn Pro Met Leu Arg Leu Cys Leu Val Lys Leu Ser
675 680 685
Val Lys Gly Ile Ala Arg Ile Gly Glu Val Ser Pro Ser Lys
690 695 700
<210> 2
<211> 2109
<212> DNA
<213> Bipolaris sorokiniana
<400> 2
atggagcagc tttcatatca gtccaaactg atttgctccg aggagtctcg acagactgga 60
tgcttcacga cactccctat ccgtattcat cctcgagatg acttagctga tgcagcaagt 120
cgtcgttttg tacaagactg ggccagagag atgagagatg gccgagaaca gtctactcat 180
ttctcattct caccagtggg aaactggtcg tccttgatct atcctgaagc gatccctgag 240
aggttggggg tcttggcgta tctctctgac ttgggcttga ttcatgatga cggtggtgaa 300
ggcttaagca ttgaagacgc gcaagccgaa catggtgaac tctgcgctgc tttagatcct 360
agtgacatca gcagcgttgc gccagattca agggcgatga aaactaagaa gcttgtctcc 420
cagtgtatgc ttgagtgtat tagccttgac cgagaattgg gcctgaagat gctcgcggcg 480
ttccgcgatg tctggctcgc tattagcgag aggaacagcg ataaggaagc gcagactatg 540
gaagaatatt tgaaatatcg aagcgacaat ggtggaatgc tagtcttctg gccgatgctg 600
cagttcagcc tcggaatgtc catatcggaa gcagaagaag cgctcgtaca gccaattatt 660
gacgcagcaa ctgaaggact tctcctagca aatgattatt tcagctggga acgcgagtat 720
agagaactcc aatccggtca atcaaagagg attgtcagtg ctgttgacct gttcatccgg 780
acgaagggac tctcaatcga ccatgcaaaa gaggaggtca aacgcatgat catcgccgct 840
gagcgcgatt tctgtcagcg tcgtgatgat ctttacaaga atcatcctga catatcattg 900
aagttgaaac gttggatcga ctgcgctggg ctagctgtct caggaaatca ctactggtgc 960
tcggcttgcc ccagacagaa tgcgtggaag gatatgactt cacagagtca taacggggct 1020
aaaagaaaaa catcacatgg cgcaacgatc agtatgcagg aagcaccctt caagaagaga 1080
aaagactcga gcttctttgg ttctcagccc agcgatgatg agccttcact ctcagaggtg 1140
tcaagctacc cattttacaa accgtcggga cttgcactgg aggcaccctc gaagtacgtc 1200
tccaacatgc cttcaaaagg cgtccggagc acgctcatag aagctctcaa cacatggctc 1260
cacgtgccgt ctgaacggct agactcgatc atgtctgtca tcaacacttt gcataatgct 1320
tcacttatcc ttgacgacct ggaagataat tcaccactac gaagaggcta cccatcgaca 1380
catatcctat ttgggcagtc tcaatcaatc aatgccgcga actttatgtt cgtacgcgct 1440
gtgcaggaag tcgcgcaaaa tctaagccca aatgccttgg ttgcagtcct tgaagaactc 1500
gagggcctgt atctaggcca gagctgggac ctatactgga aacacaacct cgcgtgtcca 1560
agtgaagccg agtacgtcaa tatgatcgac cacaagaccg gtggcatgtt tcgcatgttg 1620
ctccggatca tgcaggcaga gagcgaggtg acaccgcagc ctgattttca cagactaaca 1680
ctattgttcg gccgattttt tcagattcgt gatgactaca tgaacttcca agattacaca 1740
gcacagaaag ggctttgcga ggatcttgac gaagggaaat tttcgtaccc agttgtctac 1800
tgcttggaaa atcatccgga ataccgtggc cattttcttg gtatattccg gcaacgccct 1860
acggttgcga ccgtcaacgc atgtcccttg tcgagagaga gcaaacagca tctgactgct 1920
tgcctgaaaa agagtggagc gttcgacaaa acgatcgctt gcctgatgga tatggaacgc 1980
gacttggaat tcgagattaa ccggcttgag caacaaacgg gtgagacgaa cccaatgctg 2040
cggttgtgcc tcgtaaagct cagcgtcaaa ggaattgcga ggattggtga ggtaagccct 2100
agcaaataa 2109
<210> 3
<211> 702
<212> PRT
<213> Artificial Sequence
<400> 3
Met Glu Gln Leu Ser Tyr Gln Ser Lys Leu Ile Cys Ser Glu Glu Ser
1 5 10 15
Arg Gln Thr Gly Cys Phe Thr Thr Leu Pro Ile Arg Ile His Pro Arg
20 25 30
Asp Asp Leu Ala Asp Ala Ala Ser Arg Arg Phe Val Gln Asp Trp Ala
35 40 45
Arg Glu Met Arg Asp Gly Arg Glu Gln Ser Thr His Phe Ser Phe Ser
50 55 60
Pro Val Gly Asn Trp Ser Ser Leu Ile Tyr Pro Glu Ala Ile Pro Glu
65 70 75 80
Arg Leu Gly Val Leu Ala Tyr Leu Leu Asp Leu Gly Leu Ile His Asp
85 90 95
Asp Gly Gly Glu Gly Leu Ser Ile Glu Asp Ala Gln Ala Glu His Gly
100 105 110
Glu Leu Cys Ala Ala Leu Asp Pro Ser Asp Ile Ser Ser Val Ala Pro
115 120 125
Asp Ser Arg Ala Met Lys Thr Lys Lys Leu Val Ser Gln Cys Met Leu
130 135 140
Glu Cys Ile Ser Leu Asp Arg Glu Leu Gly Leu Lys Met Leu Ala Ala
145 150 155 160
Phe Arg Asp Val Trp Leu Ala Ile Ser Glu Arg Asn Ser Asp Lys Glu
165 170 175
Ala Gln Thr Met Glu Glu Tyr Leu Lys Tyr Arg Ser Asp Asn Gly Gly
180 185 190
Met Leu Val Phe Trp Pro Met Leu Gln Phe Ser Leu Gly Met Ser Ile
195 200 205
Ser Glu Ala Glu Glu Ala Leu Val Gln Pro Ile Ile Asp Ala Ala Thr
210 215 220
Glu Gly Leu Leu Leu Ala Asn Asp Tyr Phe Ser Trp Glu Arg Glu Tyr
225 230 235 240
Arg Glu Leu Gln Ser Gly Gln Ser Lys Arg Ile Val Ser Ala Val Asp
245 250 255
Leu Phe Ile Arg Thr Lys Gly Leu Ser Ile Asp His Ala Lys Glu Glu
260 265 270
Val Lys Arg Met Ile Ile Ala Ala Glu Arg Asp Phe Cys Gln Arg Arg
275 280 285
Asp Asp Leu Tyr Lys Asn His Pro Asp Ile Ser Leu Lys Leu Lys Arg
290 295 300
Trp Ile Asp Cys Ala Gly Leu Ala Val Ser Gly Asn His Tyr Trp Cys
305 310 315 320
Ser Ala Cys Pro Arg Gln Asn Ala Trp Lys Asp Met Thr Ser Gln Ser
325 330 335
His Asn Gly Ala Lys Arg Lys Thr Ser His Gly Ala Thr Ile Ser Met
340 345 350
Gln Glu Ala Pro Phe Lys Lys Arg Lys Asp Ser Ser Phe Phe Gly Ser
355 360 365
Gln Pro Ser Asp Asp Glu Pro Ser Leu Ser Glu Val Ser Ser Tyr Pro
370 375 380
Phe Tyr Lys Pro Ser Gly Leu Ala Leu Glu Ala Pro Ser Lys Tyr Val
385 390 395 400
Ser Asn Met Pro Ser Lys Gly Val Arg Ser Thr Leu Ile Glu Ala Leu
405 410 415
Asn Thr Trp Leu His Val Pro Ser Glu Arg Leu Asp Ser Ile Met Ser
420 425 430
Val Ile Asn Thr Leu His Asn Ala Ser Leu Ile Leu Asp Asp Leu Glu
435 440 445
Asp Asn Ser Pro Leu Arg Arg Gly Tyr Pro Ser Thr His Ile Leu Phe
450 455 460
Gly Gln Ser Gln Ser Ile Asn Ala Ala Asn Phe Met Phe Val Arg Ala
465 470 475 480
Val Gln Glu Val Ala Gln Asn Leu Ser Pro Asn Ala Leu Val Ala Val
485 490 495
Leu Glu Glu Leu Glu Gly Leu Tyr Leu Gly Gln Ser Trp Asp Leu Tyr
500 505 510
Trp Lys His Asn Leu Ala Cys Pro Ser Glu Ala Glu Tyr Val Asn Met
515 520 525
Ile Asp His Lys Thr Gly Gly Met Phe Arg Met Leu Leu Arg Ile Met
530 535 540
Gln Ala Glu Ser Glu Val Thr Pro Gln Pro Asp Phe His Arg Leu Thr
545 550 555 560
Leu Leu Phe Gly Arg Phe Phe Gln Ile Arg Asp Asp Tyr Met Asn Phe
565 570 575
Gln Asp Tyr Thr Ala Gln Lys Gly Leu Cys Glu Asp Leu Asp Glu Gly
580 585 590
Lys Phe Ser Tyr Pro Val Val Tyr Cys Leu Glu Asn His Pro Glu Tyr
595 600 605
Arg Gly His Phe Leu Gly Ile Phe Arg Gln Arg Pro Thr Val Ala Thr
610 615 620
Val Asn Ala Cys Pro Leu Ser Arg Glu Ser Lys Gln His Leu Thr Ala
625 630 635 640
Cys Leu Lys Lys Ser Gly Ala Phe Asp Lys Thr Ile Ala Cys Leu Met
645 650 655
Asp Met Glu Arg Asp Leu Glu Phe Glu Ile Asn Arg Leu Glu Gln Gln
660 665 670
Thr Gly Glu Thr Asn Pro Met Leu Arg Leu Cys Leu Val Lys Leu Ser
675 680 685
Val Lys Gly Ile Ala Arg Ile Gly Glu Val Ser Pro Ser Lys
690 695 700
<210> 4
<211> 2109
<212> DNA
<213> Artificial Sequence
<400> 4
atggagcagc tttcatatca gtccaaactg atttgctccg aggagtctcg acagactgga 60
tgcttcacga cactccctat ccgtattcat cctcgagatg acttagctga tgcagcaagt 120
cgtcgttttg tacaagactg ggccagagag atgagagatg gccgagaaca gtctactcat 180
ttctcattct caccagtggg aaactggtcg tccttgatct atcctgaagc gatccctgag 240
aggttggggg tcttggcgta tctccttgac ttgggcttga ttcatgatga cggtggtgaa 300
ggcttaagca ttgaagacgc gcaagccgaa catggtgaac tctgcgctgc tttagatcct 360
agtgacatca gcagcgttgc gccagattca agggcgatga aaactaagaa gcttgtctcc 420
cagtgtatgc ttgagtgtat tagccttgac cgagaattgg gcctgaagat gctcgcggcg 480
ttccgcgatg tctggctcgc tattagcgag aggaacagcg ataaggaagc gcagactatg 540
gaagaatatt tgaaatatcg aagcgacaat ggtggaatgc tagtcttctg gccgatgctg 600
cagttcagcc tcggaatgtc catatcggaa gcagaagaag cgctcgtaca gccaattatt 660
gacgcagcaa ctgaaggact tctcctagca aatgattatt tcagctggga acgcgagtat 720
agagaactcc aatccggtca atcaaagagg attgtcagtg ctgttgacct gttcatccgg 780
acgaagggac tctcaatcga ccatgcaaaa gaggaggtca aacgcatgat catcgccgct 840
gagcgcgatt tctgtcagcg tcgtgatgat ctttacaaga atcatcctga catatcattg 900
aagttgaaac gttggatcga ctgcgctggg ctagctgtct caggaaatca ctactggtgc 960
tcggcttgcc ccagacagaa tgcgtggaag gatatgactt cacagagtca taacggggct 1020
aaaagaaaaa catcacatgg cgcaacgatc agtatgcagg aagcaccctt caagaagaga 1080
aaagactcga gcttctttgg ttctcagccc agcgatgatg agccttcact ctcagaggtg 1140
tcaagctacc cattttacaa accgtcggga cttgcactgg aggcaccctc gaagtacgtc 1200
tccaacatgc cttcaaaagg cgtccggagc acgctcatag aagctctcaa cacatggctc 1260
cacgtgccgt ctgaacggct agactcgatc atgtctgtca tcaacacttt gcataatgct 1320
tcacttatcc ttgacgacct ggaagataat tcaccactac gaagaggcta cccatcgaca 1380
catatcctat ttgggcagtc tcaatcaatc aatgccgcga actttatgtt cgtacgcgct 1440
gtgcaggaag tcgcgcaaaa tctaagccca aatgccttgg ttgcagtcct tgaagaactc 1500
gagggcctgt atctaggcca gagctgggac ctatactgga aacacaacct cgcgtgtcca 1560
agtgaagccg agtacgtcaa tatgatcgac cacaagaccg gtggcatgtt tcgcatgttg 1620
ctccggatca tgcaggcaga gagcgaggtg acaccgcagc ctgattttca cagactaaca 1680
ctattgttcg gccgattttt tcagattcgt gatgactaca tgaacttcca agattacaca 1740
gcacagaaag ggctttgcga ggatcttgac gaagggaaat tttcgtaccc agttgtctac 1800
tgcttggaaa atcatccgga ataccgtggc cattttcttg gtatattccg gcaacgccct 1860
acggttgcga ccgtcaacgc atgtcccttg tcgagagaga gcaaacagca tctgactgct 1920
tgcctgaaaa agagtggagc gttcgacaaa acgatcgctt gcctgatgga tatggaacgc 1980
gacttggaat tcgagattaa ccggcttgag caacaaacgg gtgagacgaa cccaatgctg 2040
cggttgtgcc tcgtaaagct cagcgtcaaa ggaattgcga ggattggtga ggtaagccct 2100
agcaaataa 2109
Claims (10)
1.一种双功能萜合酶，其特征在于，其氨基酸序列如SEQ ID NO.1所示。
2.一种双功能萜合酶突变体，其特征在于，其氨基酸序列如SEQ ID NO.3所示。
3.一种分离的核酸，其特征在于，所述核酸编码权利要求1所述双功能萜合酶，所述分离的核酸的核苷酸序列如SEQ ID NO.2所示。
4.一种分离的核酸，其特征在于，所述核酸编码权利要求2所述双功能萜合酶突变体，所述分离的核酸的核苷酸序列如SEQ ID NO.4所示。
5.一种重组表达载体，其特征在于，所述重组表达载体包含编码权利要求1所述双功能萜合酶的核酸。
6.一种重组表达载体，其特征在于，所述重组表达载体包含编码权利要求2所述双功能萜合酶突变体的核酸。
7.一种重组表达转化体，其特征在于，所述重组表达转化体包含权利要求5所述的重组表达载体。
8.一种重组表达转化体，其特征在于，所述重组表达转化体包含权利要求6所述的重组表达载体。
9.双功能萜合酶突变体的催化产物，其特征在于，双功能萜合酶突变体的催化产物2和3为含5-15-双环二倍半萜骨架的化合物，其分子式为C25H40，结构式分别如式2、式3所示；双功能萜合酶突变体的催化产物4，即化合物4为含5-6-8-5-四环二倍半萜骨架的化合物，双功能萜合酶突变体的催化产物5，即化合物5为含5-6-7-3-5-五环二倍半萜骨架的化合物，二者分子式均为C25H42O，结构式分别如式4、式5所示；
10.根据权利要求9所述的双功能萜合酶突变体的催化产物，其特征在于，双功能萜合酶突变体的催化产物是：双功能萜合酶突变体在酿酒酵母S.cerevisiae BJ5464中体内酶催化反应之后，提取的突变体催化产物。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210554944.8A CN115404229A (zh) | 2022-05-19 | 2022-05-19 | 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210554944.8A CN115404229A (zh) | 2022-05-19 | 2022-05-19 | 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115404229A true CN115404229A (zh) | 2022-11-29 |
Family
ID=84157647
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210554944.8A Pending CN115404229A (zh) | 2022-05-19 | 2022-05-19 | 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115404229A (zh) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110272345A (zh) * | 2019-06-27 | 2019-09-24 | 华东理工大学 | 一类来源于植物病原真菌的5-15环二倍半萜化合物及其制备方法与应用 |
| CN110343618A (zh) * | 2019-06-27 | 2019-10-18 | 华东理工大学 | 一类helminthosporol型倍半萜类化合物及其制备方法与应用 |
| CN113046332A (zh) * | 2021-03-29 | 2021-06-29 | 华东理工大学 | 一类二倍半萜骨架化合物及其合成基因及制备方法 |
| CN113402357A (zh) * | 2021-06-25 | 2021-09-17 | 华东理工大学 | 一类5-12-5三环二倍半萜骨架化合物及其制备 |
| CN113480660A (zh) * | 2021-06-03 | 2021-10-08 | 武汉大学 | 一种嵌合萜类合酶及其应用 |
-
2022
- 2022-05-19 CN CN202210554944.8A patent/CN115404229A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110272345A (zh) * | 2019-06-27 | 2019-09-24 | 华东理工大学 | 一类来源于植物病原真菌的5-15环二倍半萜化合物及其制备方法与应用 |
| CN110343618A (zh) * | 2019-06-27 | 2019-10-18 | 华东理工大学 | 一类helminthosporol型倍半萜类化合物及其制备方法与应用 |
| CN113046332A (zh) * | 2021-03-29 | 2021-06-29 | 华东理工大学 | 一类二倍半萜骨架化合物及其合成基因及制备方法 |
| CN113480660A (zh) * | 2021-06-03 | 2021-10-08 | 武汉大学 | 一种嵌合萜类合酶及其应用 |
| CN113402357A (zh) * | 2021-06-25 | 2021-09-17 | 华东理工大学 | 一类5-12-5三环二倍半萜骨架化合物及其制备 |
Non-Patent Citations (4)
| Title |
|---|
| LAN JIANG 等: "Genome-guided investigation of anti-inflammatory sesterterpenoids with 5-15 trans-fused ring system from phytopathogenic fungi", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 105, pages 5407, XP037510745, DOI: 10.1007/s00253-021-11192-3 * |
| LAN JIANG, 等: "Genome-Based Discovery of Enantiomeric Pentacyclic Sesterterpenes Catalyzed by Fungal Bifunctional Terpene Synthases", ORGANIC LETTERS, pages 4645 - 4650 * |
| WANG, H.: "GenBank: KAF5853325.1", GENBANK * |
| 李碧霄: "两株特殊生境微生物次级代谢产物及倍半萜合酶的研究", 中国优秀硕士学位论文全文数据库 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kim et al. | Cloning and heterologous expression of the cyclooctatin biosynthetic gene cluster afford a diterpene cyclase and two p450 hydroxylases | |
| Sadre et al. | Metabolite diversity in alkaloid biosynthesis: a multilane (diastereomer) highway for camptothecin synthesis in Camptotheca acuminata | |
| CN112142585B (zh) | Mangicols二倍半萜化合物、合成方法、基因簇、核酸分子、构建体及其应用 | |
| CN106635853B (zh) | 产甘草次酸的重组酿酒酵母、其构建方法以及用途 | |
| Jiang et al. | Isolation, structure elucidation and biosynthesis of benzo [b] fluorene nenestatin A from deep-sea derived Micromonospora echinospora SCSIO 04089 | |
| CN106906201B (zh) | 一种生产橙花叔醇的萜类合酶及其应用 | |
| Toyomasu et al. | Biosynthetic gene-based secondary metabolite screening: a new diterpene, methyl phomopsenonate, from the fungus Phomopsis amygdali | |
| CN113046332B (zh) | 一类二倍半萜骨架化合物及其合成基因及制备方法 | |
| CN113186183B (zh) | 双功能二倍半萜/二萜合酶LcTPS2、编码基因及其产物和应用 | |
| CN115197172B (zh) | 二倍半萜化合物、其合成基因簇与合成方法 | |
| CN112592843B (zh) | 一种生产α-蛇麻烯的重组解脂耶氏酵母菌及其构建方法和应用 | |
| Petříčková et al. | Biosynthesis of Colabomycin E, a New Manumycin‐Family Metabolite, Involves an Unusual Chain‐Length Factor | |
| CN108239631B (zh) | 一种萜类合酶及其用途 | |
| Jiang et al. | Schultriene and nigtetraene: Two sesterterpenes characterized from pathogenetic fungi via genome mining approach | |
| Luo et al. | Characterization of a sesquiterpene cyclase from the glandular trichomes of Leucosceptrum canum for sole production of cedrol in Escherichia coli and Nicotiana benthamiana | |
| Sobreira et al. | Endophytic fungus Pseudofusicoccum stromaticum produces cyclopeptides and plant-related bioactive rotenoids | |
| Liu et al. | Genetic dereplication driven discovery of a tricinoloniol acid biosynthetic pathway in Trichoderma hypoxylon | |
| CN113652443B (zh) | 菌株及其发酵生产麦角生物碱的应用 | |
| Chen et al. | A terpene synthase-cytochrome P450 cluster in Dictyostelium discoideum produces a novel trisnorsesquiterpene | |
| Arens et al. | Exploration of biosynthetic access to the shared precursor of the fusicoccane diterpenoid family | |
| Li et al. | Biosynthetic characterization of the antifungal fernane-type triterpenoid polytolypin for generation of new analogues via combinatorial biosynthesis | |
| Shinozaki et al. | Dammaradiene synthase, a squalene cyclase, from Dryopteris crassirhizoma Nakai | |
| CN115404229A (zh) | 双功能萜合酶及其突变体以及催化产物5-15环系二倍半萜类化合物 | |
| Jiang et al. | Genome-guided investigation of anti-inflammatory sesterterpenoids with 5-15 trans-fused ring system from phytopathogenic fungi | |
| CN110317765B (zh) | 一种高产香叶醇葡萄糖苷的大肠杆菌表达菌株及其应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |